The aminoglycoside antibiotic gentamicin is able to alter metabolic activity and morphology of MDCK-C11 cells: a cell model of intercalated cells

It is well known that the aminoglycoside antibiotic gentamicin is capable of causing damage to kidney cells. Given the known involvement of Ca2+ in the nephrotoxic action of gentamicin, the purpose of this study was to establish a relationship between the concentration of intracellular Ca2+ ([Ca2+]i) and cellular cytotoxicity using MDCK-C11 cells, a clone that has several properties that resemble those of intercalated cells of the distal nephron. Changes in [Ca2+]i was determined using fluorescence microscopy. Cell viability was evaluated by the neutral red method, and cell cytotoxicity by the MTT method. The [Ca2+]i gradually increased when cells were exposed to 0.1 mM gentamicin for 10, 20, and 30 min. The presence of extracellular Ca2+ was found to be necessary to stimulate the increase in [Ca2+]i induced by gentamicin, since this stimulus disappeared by using 1.8 mM EGTA (a Ca2+ chelator). Morphological changes were observed with scanning electron microscopy in epithelial cells exposed to the antibiotic. Furthermore, with the MTT method, a decrease in metabolic activity induced by gentamicin was observed, which indicates a cytotoxic effect. In conclusion, gentamicin was able to alter [Ca2+]i, change the morphology of MDCK-C11 cells, and promote cytotoxicity.

Effects of immunosuppression with tacrolimus and mycophenolate mofetil on renal histology and function in single kidney rats submitted to ischemia and reperfusion

PURPOSE: To evaluate renal histological changes and renal function in single kidney rats submitted to renal ischemia-reperfusion and to immunosuppression with tacrolimus and mycophenolate-mofetil. METHODS: Experimental study with 80 Wistar rats distributed into control, Sham and six other groups treated with immunosuppressive drugs. Animals undergoing surgery, right nephrectomy and left renal clamping, killed on the 14th day and analyzed for renal histology, urea and creatinine. RESULTS: The group receiving tacrolimus at higher doses (T3) showed renal histological lesions indicative of early nephrotoxicity, and significant increase in urea and creatinine. The group M (mycophenolate-mofetil alone) and the group M2 (mycophenolate-mofetil combined with half the usual dose of tacrolimus) presented a slight rise in serum urea. The groups using mycophenolate-mofetil alone or combined with tacrolimus showed creatinine levels similar to that of the group T3. CONCLUSIONS: Histologically, the association of injury by ischemia-reperfusion with the use of tacrolimus or mycophenolate-mofetil alone demonstrated a higher rate of renal changes typical of early nephrotoxicity. In laboratory, the combination of injury by ischemia-reperfusion with tacrolimus at higher doses proved to be nephrotoxic. .

Effect of D-alpha-tocopherol on tubular nephron acidification by rats with induced diabetes mellitus

The objective of the present study was to determine if treatment of diabetic rats with D-alpha-tocopherol could prevent the changes in glomerular and tubular function commonly observed in this disease. Sixty male Wistar rats divided into four groups were studied: control (C), control treated with D-alpha-tocopherol (C + T), diabetic (D), and diabetic treated with D-alpha-tocopherol (D + T). Treatment with D-alpha-tocopherol (40 mg/kg every other day, ip) was started three days after diabetes induction with streptozotocin (60 mg/kg, ip). Renal function studies and microperfusion measurements were performed 30 days after diabetes induction and the kidneys were removed for morphometric analyses. Data are reported as means ± SEM. Glomerular filtration rate increased in D rats but decreased in D + T rats (C: 6.43 ± 0.21; D: 7.74 ± 0.45; D + T: 3.86 ± 0.18 ml min-1 kg-1). Alterations of tubular acidification observed in bicarbonate absorption flux (JHCO3) and in acidification half-time (t/2) in group D were reversed in group D + T (JHCO3, C: 2.30 ± 0.10; D: 3.28 ± 0.22; D + T: 1.87 ± 0.08 nmol cm-2 s-1; t/2, C: 4.75 ± 0.20; D: 3.52 ± 0.15; D + T: 5.92 ± 0.19 s). Glomerular area was significantly increased in D, while D + T rats exhibited values similar to C, suggesting that the vitamin prevented the hypertrophic effect of hyperglycemia (C: 8334.21 ± 112.05; D: 10,217.55 ± 100.66; D + T: 8478.21 ± 119.81æm²). These results suggest that D-alpha-tocopherol is able to protect rats, at least in part, from the harmful effects of diabetes on renal function.

A study of peripheral nerve function in neonates and infants.

Motor and sensory nerve conduction velocity, H-reflex and F-response have been studied in the age group showing maximum changes i.e. neonates and infants. The nerve conduction velocity in upper and lower limbs was 25 M/S and 23.75 M/S respectively in neonate age group; 34.4 M/S and 32.4 M/S respectively in infant group. A significant relationship of age with nerve conduction parameters (velocity, terminal latency) has been observed in infants group but not so in neonate group. H-reflex (late response) was elicited in both Abductor Pollicis Brevis and Soleus. It was present in small muscles of hand (i.e. APB) in all the neonates and 55% of the infants only. This could be attributed to immaturity of nervous system. However, in the lower limb, H-reflex could be elicited in 100% of infants and neonates. In the present study, the relationship of age and height with different nerve conduction parameters as well as H-reflex (latency) has been highlighted.

Estructura molecular de los receptores de diuréticos luminales / Molecular structure of luminal diuretic receptors

El riñon juega un papel central en el mantenimiento del volumen y la composición del líquido corporal. En condiciones fisiológicas, la ingesta de sal y agua, día con día, es más o menos constante, por lo que la excreción urinaria de sodio es determinante en el mantenimiento del líquido corporal. esta función la lleva a cabo el riñon gracias a la integración de la ultrafiltración, en el glomérulo, y de la reabsorción tubular a lo largo de la nefrona. El control fino de la excreción urinaria de sodio se lleva a cabo gracias a la reabsorción tubular en la nefrona distal. En este sitio, con base en el requerimiento de iones (CI- y K+) y la sensibilidad a diuréticos, tres diferentes mecanismos de transporte de sodio se han identificado en la membrana apical: 1) los cotransportadores de Na+:K+:2CI- y Na+:CI- sensibles a derivados del ácido sulfamoílbenzoico (furosemide o bumetanida) en el asa ascendente de Henle; 2) el contrasportador de Na+:CI- sensible a la benzotiadiazina (o tiazidas) en el tíbulo distal; y 3) los canales de Na+ sensibles a amilorida en el túbulo colector. La inhibición de esta proteínas con diuréticos aumenta la excreción urinaria de Na+. Gracias a técnicas avanzadas en biología molecular, como la clonación por expresión funcional en ovocitos de Xenopus laevis, se ha identificado actualmente DNAc que codifica para miembros de las familias de estos receptores. En este trabajo se presenta una revisión sobre los avances en el conocimiento de la estructura primaria y de la relación entre la estructura y función de los receptores para diuréticos. Se discuten también las posibles consecuencias que estos hallazgos tendrán para el entendimiento en la fisiología y la fisiopatología del manejo renal del sodio